End-label, free-solution capillary electrophoresis of highly charged oligosaccharides

1995 ◽  
Vol 67 (22) ◽  
pp. 4205-4209 ◽  
Author(s):  
Jan. Sudor ◽  
Milos V. Novotny
Keyword(s):  
Capillary Electrophoresis ◽  
Free Solution ◽  
Label Free

Molar Mass Profiling of Synthetic Polymers by Free-Solution Capillary Electrophoresis of DNA−Polymer Conjugates

2001 ◽  
Vol 73 (8) ◽  
pp. 1795-1803 ◽  
Author(s):  
Wyatt N. Vreeland ◽  
Claude Desruisseaux ◽  
Achim E. Karger ◽  
Guy Drouin ◽  
Gary W. Slater ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Molar Mass ◽  
Synthetic Polymers ◽  
Free Solution ◽  
Polymer Conjugates

scholarly journals Unlabeled lysophosphatidic acid receptor binding in free solution as determined by a compensated interferometric reader

2020 ◽  
Vol 61 (8) ◽  
pp. 1244-1251 ◽  
Author(s):  
Manisha Ray ◽  
Kazufumi Nagai ◽  
Yasuyuki Kihara ◽  
Amanda Kussrow ◽  
Michael N. Kammer ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Lysophosphatidic Acid ◽  
Specific Binding ◽  
Test Case ◽  
Free Solution ◽  
Label Free ◽  
Adhesive Properties ◽  
Lpa Receptor ◽  
System A ◽  
G Protein Coupled

Native interactions between lysophospholipids (LPs) and their cognate LP receptors are difficult to measure because of lipophilicity and/or the adhesive properties of lipids, which contribute to high levels of nonspecific binding in cell membrane preparations. Here, we report development of a free-solution assay (FSA) where label-free LPs bind to their cognate G protein-coupled receptors (GPCRs), combined with a recently reported compensated interferometric reader (CIR) to quantify native binding interactions between receptors and ligands. As a test case, the binding parameters between lysophosphatidic acid (LPA) receptor 1 (LPA1; one of six cognate LPA GPCRs) and LPA were determined. FSA-CIR detected specific binding through the simultaneous real-time comparison of bound versus unbound species by measuring the change in the solution dipole moment produced by binding-induced conformational and/or hydration changes. FSA-CIR identified KD values for chemically distinct LPA species binding to human LPA1 and required only a few nanograms of protein: 1-oleoyl (18:1; KD = 2.08 ± 1.32 nM), 1-linoleoyl (18:2; KD = 2.83 ± 1.64 nM), 1-arachidonoyl (20:4; KD = 2.59 ± 0.481 nM), and 1-palmitoyl (16:0; KD = 1.69 ± 0.1 nM) LPA. These KD values compared favorably to those obtained using the previous generation back-scattering interferometry system, a chip-based technique with low-throughput and temperature sensitivity. In conclusion, FSA-CIR offers a new increased-throughput approach to assess quantitatively label-free lipid ligand-receptor binding, including nonactivating antagonist binding, under near-native conditions.

scholarly journals Reply to Varma: Elucidation of the signal origin for label-free, free-solution interactions

2016 ◽  
Vol 113 (34) ◽  
pp. E4931-E4932 ◽  
Author(s):  
Darryl J. Bornhop ◽  
Michael N. Kammer ◽  
Amanda Kussrow ◽  
Robert A. Flowers
Keyword(s):  
Free Solution ◽  
Label Free

Free-Solution, Label-Free Molecular Interactions Studied by Back-Scattering Interferometry

Science ◽  
2007 ◽  
Vol 317 (5845) ◽  
pp. 1732-1736 ◽  
Author(s):  
D. J. Bornhop ◽  
J. C. Latham ◽  
A. Kussrow ◽  
D. A. Markov ◽  
R. D. Jones ◽  
...  
Keyword(s):  
Molecular Interactions ◽  
Free Solution ◽  
Label Free ◽  
Back Scattering
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